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Plant and Cell Physiology Advance Access originally published online on September 17, 2007
Plant and Cell Physiology 2007 48(10):1393-1403; doi:10.1093/pcp/pcm120
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Rapid Paper

Chemical Genetic Screening Identifies a Novel Inhibitor of Parallel Alignment of Cortical Microtubules and Cellulose Microfibrils

Arata Yoneda1, Takumi Higaki2, Natsumaro Kutsuna2, Yoichi Kondo1, Hiroyuki Osada3, Seiichiro Hasezawa2 and Minami Matsui1,*

1Plant Science Center, RIKEN, 1-7-22, Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045 Japan
2Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwanoha 5-1-5, Kashiwa, Chiba, 277-8562 Japan
3Discovery Research Institute, RIKEN, 2-1, Hirosawa, Wako, Saitama, 351-0198 Japan

*Corresponding author: E-mail, minami{at}postman.riken.go.jp; Fax, +81-45-503-9586.


  Abstract

It is a well-known hypothesis that cortical microtubules control the direction of cellulose microfibril deposition, and that the parallel cellulose microfibrils determine anisotropic cell expansion and plant cell morphogenesis. However, the molecular mechanism by which cortical microtubules regulate the orientation of cellulose microfibrils is still unclear. To investigate this mechanism, chemical genetic screening was performed. From this screening, ‘SS compounds’ were identified that induced a spherical swelling phenotype in tobacco BY-2 cells. The SS compounds could be categorized into three classes: those that disrupted the cortical microtubules; those that reduced cellulose microfibril content; and thirdly those that had neither of these effects. In the last class, a chemical designated ‘cobtorin’ was found to induce the spherical swelling phenotype at the lowest concentration, suggesting strong binding activity to the putative target. Examining cellulose microfibril regeneration using taxol-treated protoplasts revealed that the cobtorin compound perturbed the parallel alignment of pre-existing cortical microtubules and nascent cellulose microfibrils. Thus, cobtorin could be a novel inhibitor and an attractive tool for further investigation of the mechanism that enables cortical microtubules to guide the parallel deposition of cellulose microfibrils.

Keywords: Cell swelling - Cellulose microfibril - Chemical genetics - Cortical microtubule - Inhibitor

Abbreviations: APM, amiprofos methyl; DCB, 2,6-dichlorobenzonitrile; GFP, green fluorescent protein

(Received August 14, 2007; Accepted September 12, 2007)
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